FIG. 1.

HCV infection enhances AR activation in the presence of DHT. (A) AR mRNA expression varies in different hepatocyte cell lines. RNAs from Huh-7, IHH, and HepG2 cells were subjected to real-time RT-PCR for AR and GAPDH gene expression. The PC-3 (AR-negative) cell line was used as a negative control. The results are presented as relative expression levels of AR normalized with GAPDH housekeeping gene as an internal control. The expression level of AR in Huh-7 cells was arbitrarily chosen as 1 for comparison. (B) HCV protein expression enhances androgen-dependent AR-responsive gene expression. IHH were cotransfected with pARE-luciferase reporter construct (pARE3-luc) and HCV-FL expression plasmid DNA or empty vector (negative control) in the presence or absence of DHT. Cell lysates were prepared after 48 h of transfection, and the luciferase activity was measured. The relative luciferase activity of the negative control in the absence of DHT was arbitrarily set as 1 for comparison. Luciferase activities are presented as an average from three independent experiments (mean ± the standard deviation [SD]). (C) Huh-7 cells were similarly used for transfection with HCV-FL. Experiments were performed as described in panel B for measurement of luciferase activity. (D) IHH were infected with cell culture-grown HCV (clone H77) for 24 h, followed by transfection with pARE3-luc. Experiments were performed as described in panel B for measurement of the luciferase activity. The lowest level of significance was P < 0.001.